Reconstitution of the Golgi apparatus after microinjection of rat liver Golgi fragments into Xenopus oocytes

Journal Article · Sat Apr 01 04:00:00 EST 1989 · J. Cell Biol.; (United States)

DOI:https://doi.org/10.1083/jcb.108.4.1257· OSTI ID:6102351

Paiement, J; Jolicoeur, M; Fazel, A; Bergeron, J J

We have studied the reconstitution of the Golgi apparatus in vivo using an heterologous membrane transplant system. Endogenous glycopeptides of rat hepatic Golgi fragments were radiolabeled in vitro with (3H)sialic acid using detergent-free conditions. The Golgi fragments consisting of dispersed vesicles and tubules with intraluminal lipoprotein-like particles were then microinjected into Xenopus oocytes and their fate studied by light (LM) and electron microscope (EM) radioautography. 3 h after microinjection, radiolabel was observed by LM radioautography over yolk platelet-free cytoplasmic regions near the injection site. EM radioautography revealed label over Golgi stacked saccules containing the hepatic marker of intraluminal lipoprotein-like particles. At 14 h after injection, LM radioautographs revealed label in the superficial cortex of the oocytes between the yolk platelets and at the oocyte surface. EM radioautography identified the labeled structures as the stacked saccules of the Golgi apparatus, the oocyte cortical granules, and the plasmalemma, indicating that a proportion of microinjected material was transferred to the surface via the secretion pathway of the oocyte. The efficiency of transport was low, however, as biochemical studies failed to show extensive secretion of radiolabel into the extracellular medium by 14 h with approximately half the microinjected radiolabeled constituents degraded. Vinblastine (50 microM) administered to oocytes led to the formation of tubulin paracrystals. Although microinjected Golgi fragments were able to effect the formation of stacked saccules in vinblastine-treated oocytes, negligible transfer of heterologous material to the oocyte surface could be detected by radioautography.

• Research Organization: Universite de Montreal, Quebec (Canada)
• OSTI ID: 6102351
• Journal Information: J. Cell Biol.; (United States), Journal Name: J. Cell Biol.; (United States) Vol. 108:4; ISSN JCLBA
• Country of Publication: United States
• Language: English

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550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMPHIBIANS
ANIMALS
AQUATIC ORGANISMS
AUTORADIOGRAPHY
BIOLOGICAL MARKERS
BIOLOGICAL PATHWAYS
BODY
CELL CONSTITUENTS
CYTOPLASM
DIGESTIVE SYSTEM
ELECTRON MICROSCOPY
FROGS
GERM CELLS
GLANDS
GLYCOPROTEINS
LABELLED COMPOUNDS
LIVER
MAMMALS
MEMBRANE TRANSPORT
MICROSCOPY
MOLECULAR WEIGHT
OOCYTES
ORGANIC COMPOUNDS
ORGANOIDS
ORGANS
PROTEINS
RATS
RODENTS
TRITIUM COMPOUNDS
VERTEBRATES